1.Establishment of SLC6A4 gene silencing stable cell line in PC12 cells and the effect of SLC6A4 gene silencing on apoptosis of hypoxia PC12 cells
Dongmei ZHANG ; Yingchun GAO ; Lei HE ; Keming CHEN ; Wenbin LI ; Rong WANG ; Huiping MA
Journal of Pharmaceutical Practice and Service 2022;40(5):389-394
Objective To construct SLC6A4-shRNA lentiviral vector, establish PC12 cells stable transformation cell line,and detect the effect of SLC6A4 gene silencing on hypoxia induced PC12 cells apoptosis. Methods Three specific targets sequence of SLC6A4 were designed and short hairpin RNA was synthesized, and then were recombined into shRNA expression vector GV248 plasmid, with non-homology shRNA sequence as negative control. The connection products were switched to competent cells. After dentification and sequencing, the vectors were co-transfected with the auxiliary vectors into 293T cells in order to produce recombinant shRNA lentiviral particles. Then, PC12 cells were infected with the recombinant lentiviral and screened by puromycin. The PC12 cells were divided into two groups: lentiviral negative control group (NC-shRNA) and SLC6A4-silenced group (SLC6A4-shRNA). The expression of SLC6A4 mRNA was detected by real-time fluorescence quantitative and the 5-HTT protein level was assayed by Western blot, The effect of SLC6A4 gene silencing on hypoxia induced apoptosis was detected by flow cytometry. Results The SLC6A4-shRNA lentiviral expression vector was constructed and the recombinant lentiviral particles by packaging the 293T cells were obtained, the stably infected PC12 cells were established after filtering. Compared with negative control group, the expression level of SLC6A4 gene and 5-HTT protein in SLC6A4-shRNA group was suppressed notably (P<0.01). It was confirmed that lentiviral vector could effectively silence SLC6A4 gene in PC12 cells and SLC6A4 gene silencing could decrease apoptosis rate of PC12 cells under hypoxia condition. Conclusion The SLC6A4 gene of PC12 cells could be effectively silenced by shRNA lentivirus vector,which could reverse hypoxia induced apoptosis.
2.Clinical trial of nifedipine controlled-release tablets on reducing blood pressure in the treatment of patients with hypertension at high altitude
Qin HUANG ; Zizhao GAO ; Zhuoma NIYANG ; Gele SUONAN ; Rong WANG
Journal of Pharmaceutical Practice and Service 2022;40(5):395-398
Objective To observe the clinical efficacy and safety of nifedipine controlled-release tablets on the antihypertensive effect of hypertensive patients under high altitude environment. Methods 42 hypertensive inpatients in the 940th hospital (altitude 1500 ) were set to the plain hypertension group, and 42 cases of hypertensive inpatients in Bayi hospital (altitude 3800 m) were set to the plateau hypertension group. Both groups of patients were given nifedipine controlled-release tablets 30 mg daily, taken orally in the morning for 6 consecutive days. Monitor blood pressure and heart rate three times a day to compare the clinical efficacy and occurrence of adverse drug reactions in the two groups. Results After treatment, the total effective rates of the high-altitude hypertension group and the plain hypertension group were 47.62% (20 cases/42 cases) and 76.19% (32 cases/42 cases) respectively with no statistical difference (P<0.05). The adverse drug reactions of the two groups of patients were tachycardia and palpitations. The incidence of total adverse drug reactions in the high-altitude hypertension group and the plain hypertension group were 14.29% and 11.90% respectively with no statistical difference (P>0.05). Conclusion The high-altitude hypoxic environment could affect the antihypertensive effect of nifedipine controlled-release tablets, which could not control the patient's blood pressure effectively in the short term.
3.A preliminary study on the mechanism of 7-HEC on high altitude cerebral edema
Zhiqun SHI ; Yingchun GAO ; Dongmei ZHANG ; Keming CHEN ; Linlin JING ; Huiping MA
Journal of Pharmaceutical Practice and Service 2022;40(5):399-402
Objective To study the possible mechanism of 7-hydroxyethyl chrysin (7-HEC) on high altitude cerebral edema (HACE). Methods A rat model of high altitude cerebral edema was established. The activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in rat brain tissues were measured. The expression levels of apoptosis, cell cycle and autophagy related proteins were detected by Western blotting to explore the protective effect of 7-HEC on high altitude cerebral edema and its mechanism. Results Compared with the control group, the content of MDA in the brain tissue of the hypoxia model group was significantly up-regulated; the activity of SOD was significantly down-regulated, the relative expression of CyclinD1, CyclinE1, CDK6 and CDK2, apoptotic proteins Bcl-2, PARP, and autophagy protein LC3-B were down-regulated; and the relative expression of apoptotic protein Bax and autophagy protein P62 were up-regulated; the difference was statistically significant (P<0.05); Compared with the hypoxia model group, the content of MDA was down-regulated and the activity of SOD was significantly up-regulated in the 7-HEC administration group. The relative expression of CyclinD1, CyclinE1, CDK6, CDK2, apoptotic proteins Bcl-2, PARP, autophagy protein LC3-B was up-regulated and the relative expression of apoptotic proteins Bax and the relative expression of autophagy protein P62 was down-regulated in the 7-HEC administration group. The difference was statistically significant (P<0.05). Conclusion 7-HEC has a certain protective effect on high altitude cerebral edema, and its mechanism may be related to the regulation of cell cycle, autophagy, apoptosis and oxidative stress pathways.
4.Protective effect of Saussurea involucrata flavone capsule on myocardial injury induced by hypoxia at high altitude in mice
Pengpeng ZHANG ; Lin LI ; ZhiQun SHI ; Lei HE ; Huiping MA
Journal of Pharmaceutical Practice and Service 2022;40(5):403-407
Objective To study the protective effect and mechanism of Saussurea involucrata flavone capsule on the myocardial tissue of mice in simulated plateau hypoxia environment. Methods 64 Balb/c mice were randomly divided into normal control group, hypoxia model group, positive control group and Xuelian flavonoid capsule group. Myocardial tissue was observed microscopically and ultra-structurally, and the changes of hypoxia-related indexes were detected. The changes in the transcription levels of hypoxia-related genes were detected by RT-PCR, and the changes in the protein expressions of hypoxia-related genes were detected by Western blotting to study the mechanism of action of Xuelian flavone capsules. Results Saussurea involucrata flavone capsule had significantly alleviated the pathological damage to the myocardium of mice caused by simulated altitude hypoxia, increased the activity of T-AOC in myocardial tissue, reduced the accumulation of LD, and also reduced the expression levels of HIF-1α and VEGF mRNA in myocardial tissue, increased the SOD, CAT mRNA and protein expression levels. Conclusion Saussurea involucrata flavone capsule have good anti-altitude hypoxia effect, which could protect myocardial tissue structure and function, regulate energy metabolism, and improve antioxidant capacity. The mechanism might be related to improving the antioxidant capacity, regulating energy metabolism, and affecting the protein expression of HIF-1α, VEGF, SOD and CAT mRNA in high altitude hypoxic mice.
5.Research progress of Tibetan medicine against plateau hypoxia
Zihan WANG ; Anpeng ZHAO ; Hongfang MU ; Qianwen GUO ; Junfei CHENG ; Rong WANG
Journal of Pharmaceutical Practice and Service 2022;40(5):408-415
Tibetan is a kind of medicine which extensively absorbed and fused the Chinese Traditional Medicine, Indian medicine and food through long-term practice and form the unique medical system. At the same time, the Qinghai Tibet plateau has characteristics of high altitude and lack of resources and oxygen, which has formed its unique geographical environment and grow a lot of rare medicinal plants. Tibetan medicine has a long history and is one of the relatively complete and influential ethnic medicines in China, which has played an important role in the treatment of plateau diseases. In recent years, with the increasing of economic activities in the plateau, the plateau anoxia had greatly affected the working capacity of the plateau. According to "Chinese medicine" through the literature, the pharmacological activities, active ingredients, growing environment of Tibetan medicine with good plateau hypoxia effect were reviewed and according to the site, which could be divided into the lung protection medicine, the cardiac protection medicine, the brain protection medicine, the liver protection medicine, the improvement of fatigue medicine and other medicine, which could provide theoretical basis for finding more potential anti-hypoxia drugs.
6.Research progress of atorvastatin gene polymorphism
Qin HUANG ; Gele SUONAN ; Wenbin LI ; Yuemei SUN ; Rong WANG
Journal of Pharmaceutical Practice and Service 2022;40(5):416-421
Atorvastatin is a blood lipid-lowering drug widely used clinically. Long-term use can prevent and reduce the occurrence of atherosclerotic cardiovascular disease (ASCVD). However, the efficacy of atorvastatin has significant inter-individual differences. Some individuals failed to achieve the expected lipid-lowering target value or had serious adverse reactions, which were related to the genetic diversity between individuals. Genetic variation can lead to differences in drug configuration, clinical efficacy and adverse reactions. The drug metabolism enzymes, drug transporters, drug targets and genetic polymorphisms related to lipid metabolism were reviewed in this paper that affect the drug response of atorvastatin, and from the gene level to explore the reasons for the differences in the pharmacokinetics, pharmacodynamics and susceptibility to adverse reactions of different individuals using atorvastatin.
7.Advances in antidepressant therapy related to gut microbiota
Qiannan WANG ; Xinhui HUANG ; Minxu YANG ; Xingrui YANG ; Kehan ZHU ; Tingting ZHOU
Journal of Pharmaceutical Practice and Service 2022;40(5):422-426
Objective This paper introduces the research progress on the pathogenesis of depression related to gut microbiota and provides the resources for the subsequent development of antidepressant drugs targeting gut microbiota. Methods 33 literatures on gut microbiota and depression in recent years were reviewed. The changes of gut microbiota diversity under depression were discussed from the perspectives of phylum, family and genus. The relationship between gut microbiota and the pathogenesis of depression was expounded at the molecular level, and the existing relevant studies were summarized. The feasibility of drug development targeting gut microbiota was explored. Results There is a relationship between gut microbiota disorder and depression. Existing biological agents such as probiotics can alleviate depression by adjusting the disorder of gut microbiota. Conclusion The imbalance of gut microbiota is closely related to the occurrence of depression, and the development of drugs targeting gut microbiota may become a new way to treat depression.
8.Research progress of anti-radiation natural products
Yiqing YAO ; Jiahao FANG ; Huilin MA ; Xuan WANG ; Diya LYU
Journal of Pharmaceutical Practice and Service 2022;40(5):427-432
With the rapid developments of science and technology, the diagnosis technology based on nuclear physics and radiotherapy technology are widely used in medicine, but radiation at the same time could have different levels of damage to human body. Therefore, it is of great significance to research and develop drugs that can prevent and treat radiation damage. The research progresses and prospects of radiation-resistant natural products, such as polysaccharides, flavonoids, phenolic acids, saponins and so on, were reviewed in this paper in order to provide a reference for further developments.
9.Overview and prospects of great health products of traditional Chinese medicine for anti-epidemic application
Jianyong HAN ; Yiping JIANG ; Ting HAN ; Hailiang XIN
Journal of Pharmaceutical Practice and Service 2022;40(5):433-436
Anti-epidemic traditional Chinese medicine health products are in line with the Healthy China strategy, play an important role in epidemic prevention and control, with adantages of diverse types, obvious effects, easy acceptance, and convenient administration. With rapid progess in science and technology,the research and development of new products is increasing and getting closer to the actual needs of different groups and different scenarios. With the rapid expansion of health industry of anti-epidemic traditional Chinese medicine, while bringing more products to meet the needs of the public, it also fully exposes its shortcomings. This paper reviews its application and research and development status, and proposes suggestions and countermeasures for some of the existing problems, with a view to assisting the application and research and development of traditional Chinese medicine health products industry.
10.Preparation and characterization of photosensitive ROS-responsive rapamycin liposomes
Xiqing GAO ; Guangzhao LU ; Ying LU ; Hao ZOU
Journal of Pharmaceutical Practice and Service 2022;40(5):437-441
Objective To study the preparation process and properties of photosensitive ROS (Reactive oxygen species) responsive rapamycin liposome, and to develop a stable and efficient stimulus-responsive liposome carrier. Methods Rapamycin liposomes were prepared by thin film dispersion method. The particle size and Zeta potential were determined by Malvern laser particle size analyzer. An assay method for rapamycin was established by HPLC. In vitro release characteristics of rapamycin liposomes were investigated by reverse dialysis after irradiation with near-infrared light. Results The particle size of rapamycin liposome was less than 200 nm and the PDI value was less than 0.200. Rapamycin showed a good linear relationship with peak area in the range of 0.2-40 μg/ml, with the correlation coefficient of 0.9995. Encapsulation rate of rapamycin liposomes was > 94.20%. The release efficiency of rapamycin liposomes reached 60% within 12 h after irradiation with 730 nm near infrared light for 5 min. Conclusion Photosensitive ROS-responsive rapamycin liposomes were successfully prepared, which had high encapsulation rate and stimulation response efficiency in vitro.